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One way to do experiments on gene conversion?

Transfection with heteroduplex SPP1 DNA

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Summary

Competent cells of B. subtilis were transfected with heteroduplex SPP1 DNA, made by annealing complementary strands of wild type and 21 plaque type mutant DNAs. The frequencies of cells yielding mutant and wild type, only wild type, and only mutant phages were determined by single burst analyses of transfected cells. The data obtained reveal that an effective mechanism is operating in B. subtilis, which converts heterozygous to homozygous molecules prior to their replication. This “correction” mechanism is asymmetric with regard to the strand which is preferentially corrected in a given heteroduplex pair. The direction of asymmetry thus defined depends on the marker introduced into a particular heteroduplex. The efficiency of correction varies with the markers used and is correlated to the position of markers in the genetic map. From this correlation, the direction of replication of the SPP1 genome is deduced. The frequency distribution of wild type and mutant phages in cells yielding both genotypes indicates that both strands of the input DNA contribute equally to the production of progeny, i.e. DNA replication is symmetric.

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References

  • Baas, P. D., Jansz, H. S.: Personal communication (1969).

  • Berger, H.: Genetic analysis of T4D phage heterozygotes produced in the presence of 5-fluorodeoxyuridine. Genetics 52, 729–746 (1965).

    Google Scholar 

  • Biswal, N., Kleinschmidt, A. K., Spatz, H. Ch., Trautner, T. A.: Physical properties of the DNA of bacteriophage SP5O. Molec. Gen. Genetics 100, 39–55 (1967).

    Google Scholar 

  • Bodmer, W. F.: Integration of deoxyribonuclease-treated DNA in Bacillus subtilis transformation. J. gen. Physiol. 49, part 2, 233–258 (1966).

    Google Scholar 

  • — Ganesan, A. T.: The molecular basis for recombination in Bacillus subtilis transformation. Genetics 50, 717–738 (1964).

    Google Scholar 

  • Boon, T., Zinder, N. D.: A mechanism for genetic recombination generating one parent and one recombinant. Proc. nat. Acad. Sci. (Wash.) 64, 573–577 (1969).

    Google Scholar 

  • Bresler, S. W., Kreneva, R. A., Kushev, V. V.: Correction of molecular heterozygotes in the course of transformation. Molec. Gen. Genetics 102, 257–268 (1968).

    Google Scholar 

  • Chilton, M. D.: Transforming activity in both complementary strands of Bacillus subtilis DNA. Science 157, 817–819 (1967).

    Google Scholar 

  • Darlington, A. J., Bodmer, W. F.: Events occurring at the site of integration of a DNA molecule in Bacillus subtilis transformation. Genetics 60, 681–684 (1968).

    Google Scholar 

  • Doerfler, W., Hogness, D. S.: Gene orientation in bacteriophage lambda as determined from the genetic activities of heteroduplex DNA formed in vitro. J. molec. Biol. 33, 661–678 (1968).

    Google Scholar 

  • Doerman, A. H., Boehner, L.: An experimental analysis of bacteriophage T4 heterozygotes. I. Mottled plaques from crosses involving six r II loci. Virology 21, 551–567 (1963).

    Google Scholar 

  • Emerson, S.: Quantitative implications of the DNA-repair model of gene conversion. Genetics 53, 475–485 (1966).

    Google Scholar 

  • Ephrussi-Taylor, H. E., Gray, T. C.: Genetic studies of recombining DNA in Pneumococcal transformation. J. gen. Physiol. 49, part 2, 211–231 (1966).

    Google Scholar 

  • Fox, M., Allen, M.: On the mechanism of deoxyribonucleate integration in pneumococcal transformation. Proc. nat. Acad. Sci. (Wash.) 52, 412–419 (1964).

    Google Scholar 

  • Freese, E., Bautz, E., Freese, E. B.: The chemical and mutagenic specificity of hydroxylamine. Proc. nat. Acad. Sci. (Wash.) 47, 845–855 (1961).

    Google Scholar 

  • —, Strack, H. B.: Induction of mutations in transforming DNA by hydroxylamine. Proc. nat. Acad. Sci. (Wash.) 48, 1796–1803 (1962).

    Google Scholar 

  • Green, D. M.: Intracellular inactivation of infective SP82 bacteriophage DNA. J. molec. Biol. 22, 1–13 (1966).

    Google Scholar 

  • Guerrini, F., Fox, M. S.: Effects of DNA repair in transformation-heterozygotes of pneumococcus. Proc. nat. Acad. Sci. (Wash.) 59, 1116–1123 (1968).

    Google Scholar 

  • Havender, W. R.: Recombinational and physico-chemical studies on the topography of the chromosome of Bacillus subtilis bacteriophage SP50. Thesis, Berkeley (1969).

  • Hershey, A., Chase, M.: Genetic recombination and heterozygosis in bacteriophage. Cold Spr. Harb. Symp. quant. Biol. 16, 471–479 (1951).

    Google Scholar 

  • Hoch, J. A., Barat, M., Anagnostopoulos, C.: Transformation and transduction in recombination defective mutants of Bacillus subtilis. J. Bact. 93, 1925–1937 (1967).

    Google Scholar 

  • Holliday, R., Whitehouse, H. L. K.: The wrong way to think about gene conversion? Molec. Gen. Genetics 107, 85–93 (1970).

    Google Scholar 

  • Kellenberger, G., Zichichi, M. L., Epstein, H. T.: Heterozygosis and recombination of bacteriophage λ. Virology 17, 44–55 (1962).

    Google Scholar 

  • Levinthal, C.: Recombination in phage T2: its relationship to heterozygosis and growth. Genetics 39, 169–184 (1954).

    Google Scholar 

  • Okubo, S., Romig, W. R.: Comparison of ultraviolet sensitivity of Bacillus subtilis bacteriophage SP02 and its infectious DNA. J. molec. Biol. 14, 130–142 (1965).

    Google Scholar 

  • — Impaired transformability of a Bacillus subtilis mutant sensitive to mytomycin C and ultraviolet radiation. J. molec. Biol. 15, 440–454 (1966).

    Google Scholar 

  • —, Stodolski, M., Strauss, B.: The possible role of recombination in the infection of competent Bacillus subtilis by bacteriophage deoxyribonucleic acid. Virology 24, 552–562 (1964).

    Google Scholar 

  • Pene, J. J., Romig, W. R.: On the mechanism of genetic recombination in transforming B. subtilis. J. molec. Biol. 9, 236–245 (1964).

    Google Scholar 

  • Riva, S., Posinelli, M., Falaschi, A.: A new phage of Bacillus subtilis with infectious DNA having separable strands. J. molec. Biol. 35, 347–356 (1968).

    Google Scholar 

  • Rottländer, E., Trautner, T. A.: Genetic and transfection studies with B. subtilis phage SP50. I. Phage mutants with restricted growth on B. subtilis strain 168. Molec. Gen. Genetics, 108, 47–60 (1970).

    Google Scholar 

  • Russo, V. E. A., Stahl, M. M., Stahl, F. W.: On the transfer of information from old to new chains of DNA duplexes in phage λ-destruction of heterozygotes. Proc. nat. Acad. Sci. (Wash.) 65, 363–367 (1970).

    Google Scholar 

  • Spizizen, J.: Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleate. Proc. nat. Acad. Sci (Wash.) 44, 1072–1078 (1958).

    Google Scholar 

  • Stahl, F. W.: One way to think about gene conversion. Genetics 61, Suppl. 1–13 (1969).

  • Tessman, I.: Mutagenic treatment of double- and single-stranded DNA phages T4 and S13 with hydroxylamine. Virology 35, 331–333 (1968).

    Google Scholar 

  • Tomizawa, J.-I.: Molecular mechanism of genetic recombination: Joint molecules and their conversion to recombinant molecules. J. cell. Physiol. 70, Suppl. 1, 201–213 (1967).

    Google Scholar 

  • Trautner, T. A.: Untersuchungen an Heterozygoten des Phagen T1. Z. Vererbungsl. 89, 264–271 (1958).

    Google Scholar 

  • Trautner, T. A. Spatz, H. Ch.: Transfection with heteroduplex phage DNA. Abstracts of papers presented at the 1969 European Phage Meeting.

  • Whitehouse, H. L. K.: Towards an understanding of the mechanism of heredity. 2nd ed. London: E. Arnold, Ltd. 1968.

    Google Scholar 

  • —: The mechanism of genetic recombination. Biol. Rev. 45, 265–315 (1970).

    Google Scholar 

  • — Hastings, P. J.: The analysis of genetic recombination on the polaron hybrid DNA model. Genet. Res. Camb. 6, 27–92 (1965).

    Google Scholar 

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Authors and Affiliations

  1. Abt Trautner, Max-Planck-Institut für Molekulare Genetik, Berlin

    H. Ch. Spatz & T. A. Trautner

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  1. H. Ch. Spatz
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  2. T. A. Trautner
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Communicated by P. Starlinger

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Spatz, H.C., Trautner, T.A. One way to do experiments on gene conversion?. Molec. Gen. Genetics 109, 84–106 (1970). https://doi.org/10.1007/BF00334048

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  • DOI: https://doi.org/10.1007/BF00334048

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